Note: Descriptions are shown in the official language in which they were submitted.
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GnRH Analogue Formulations
The present invention relates to formulation precursors (pre-formulations) for
the in
situ generation compositions for the controlled release of active agents such
as
GnRH agonists and/or antagonists and methods of treatnlent with such
formulations.
In particular, the invention relates to pre-formulations of amphiphilic
components
and at least one GnRH agonist and/or antagonist, or other active agent for
parenteral
application, which undergo phase transition upon exposure to aqueous fluids,
such
as body fluids, thereby forming a controlled release matrix.
r
Many bioactive agents including pharmaceuticals, nutrients, vitamins and so
forth
have a "functional window". That is to say that there is a range of
concentrations
over which these agents can be observed to provide some biological effect.
Where
the concentration in the appropriate part of the body (e.g. locally or as
demonstrated
by serum concentration) falls below a certain level, no beneficial effect can
be
attributed to the agent. Similarly, there is generally an upper concentration
level
above which no fitrther benefit is derived by increasing the concentration. In
some
cases increasing the concentration above a particular level results in
undesirable or
even dangerous effects.
Some bioactive agents have a long biological half-life and/or a wide
functional
window and thus may be administered occasionally, maintaining a functional
biological concentration over a substantial period of time (e.g. 6 hours to
several
days). In other cases the rate of clearance is high and/or the functional
window is
narrow and thus to maintain a biological concentration within this window
regular
(or even continuous) doses of a small amount are required. This can be
particularly
difficult where non-oral routes of administration (e.g. parenteral
administration) are
desirable or necessary, since self-administration may be difficult and thus
cause
inconvenience and/or poor compliance. In such cases it would be advantageous
for
a single administration to provide active agent at a therapeutic level over
the whole
period during which activity is needed.
Gonadotropin-releasing hormone (GnRH) (also known as Luteinizing hormone-
releasing hormone (LHRH) and gonadorelin) is a naturally occurring deca-
peptide
hormone which acts largely on the pituitary gland in humans. The effect of
GnRH is
release pituitary hormones such as luteinizing hormone (LH) and follicle-
stimulating
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hormone (FSH) and to contribute to the hormonal control of processes such as
ovulation. The release of LH (also known as gonadotrophin) promotes ovarian
and
testicular steroidogenesis and thus controls levels of progesterone, estrogen
and
testosterone/ dihydrotestosterone (DHT).
GnRH itself is a post-translationally modified decapeptide of structure pyro-
Glu-
His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2 (GnRH-I). Two natural varients are
also known, GNRH-II having 5-His, 7-Trp, 8-Tyr substitutions and GnRH III
having 7-Trp, 8-Leu. Several peptide analogues with agonistic properties are
known, most of whi,ch have the10-Gly-NH2 replaced with N-Et-NH2. Fer-tirelin
has
10-Gly to N-Et-NH2 substitution only, while analogues having additional
substitutions over GnRH-Iinclude Leuprorelin (Leuprolide), (6-D-Leu),
Buserelin
(6-Ser(But)), Histrelin (6-d-His(Imbzl)), Deslorelin (6-d-Trp). Another common
nona-peptide agonist is Goserelin which is substituted with 6-Ser.(Bu) and has
10-
Gly-NH2 replaced by AzaGly-NH2. Narafelin (6-d-Nal) and Triptorelin (6-d-Trp)
both retain the 10-Gly-NH2 group. The structures of the two most common GnRH
agonists (Leuprolide and Goserelin) are shown below.
Leuprolide: pyro-Glu-His-Trp-Ser-Tyr-D-Leu-Leu-Arg-Pro- N-Et-NH2 (acetate)
Goserelin: pyro-Glu-His-Trp-Ser-Tyr-D-Ser(But)-Leu-Arg-Pro-Azgly-NH2 (acetate)
A small number of GnRH antagonists are also known, again based on the GnRH-I
structure. These include Abarelix (D-Ala-D-Phe-D-Ala-Ser-Tyr-D-Asp-Leu-
Lys('Pr)-Pro-D-Ala), Antarelix (D-Nal-D-Phe-D-Pal-Ser-Phe-D-Hcit-Leu-Lys('Pr)-
Pro-D-Ala); Cetrorelix (D Nal-D-Phe-D-Pal-Ser-Tyr-D-Cit-Leu-Arg-Pro-D-Ala),
Ganirelix (D-Nal-D-Phe-D-Pal-Ser-Tyr-D-hArg-Leu-HArg-Pro-D-Ala), Itrelix (D-
Nal-D-Phe-D-Pal-Ser-NicLys-D- NicLys -Leu-Lys('Pr)-Pro-D-Ala) and Nal-Glu (D-
Nal-D-Phe-D-Pal-Ser-D-Glu-D- Glu -Leu-Arg-Pro-D-Ala).
Administration of single doses of a GnRH agonist, such as leuprolide,
stimulates
pituitary release of gonadotropins (i.e., LH and FSH), resulting in increased
serum
LH and FSH concentrations and stimulation of ovarian and testicular
steroidogenesis. Transient increases in serum testosterone and
dihydrotestosterone
(DHT) in males and in serum estrone and estradiol concentrations in
premenopausal
females are observed during initial therapy with single daily doses of the
drug.
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Although the effect of a potent GnRH agonist during short-term and/or
intermittent
therapy is stimulation of steroidogenesis, the principal effect of the drug in
animals
and humans during long-term administration is inhibition of gonadotropin
secretion
and suppression of ovarian and testicular steroidogenesis. The exact
mechanism(s)
of action has not been fully elucidated, but continuous therapy with a GnRH
agonist
apparently produces a decrease in the number of pituitary GnRH and/or
testicular
LH receptors, resulting in pituitary and/or testicular desensitization,
respectively.
The drug does not appear to affect receptor affinity for gonadotropins.
Leuprolide's
mechanism of action may also involve inhibition and/or induction of enzymes
that
control steroidogenesis. Other mechanisms of action may include secretion of
an LH
molecule with altered biologic activity or impairment of normal pulsatile
patterns of
LH and FSH secretion.
A nutnber of serious medical indications are related to and/or affected by the
concentration of gonadal steroid hormones. These include certain neoplastic
diseases, including cancers, especially of the breast and prostate, and benign
prostatic hypertrophy; premature or delayed puberty in adolescents;
hirsuitism;
alzheimer's disease; and certain conditions relating to the reproductive
system, such
as hypogonadism, anovulation, amenorrhea, oligospermia, endometriosis,
leiomyomata (uterine fibroids), premenstrual syndrome, and polycystic ovarian
disease. Control of this system is also important in in vitro fertilisation
methods.
Although treatment with a GnRH agonist might be expected to exacerbate
conditions affected by gonadal steroid hormone concentration, the down-
regulation
effect discussed above results in the decrease of these hormones to castrate
level if
therapy is continued for around 2 weeks or longer. As a result, hormone-
receptive
tumours such as certain prostate and breast cancer, as well as precoucious
puberty
and many of the other conditions mentioned above can be improved or palliated
by
long-term GnRH agonist therapy.
Evidently, treatments involving long-term, ongoing therapy, especially where
intermittent dosing could provide a reversal of the effect desired, require
careful
monitoring and patient compliance in order to ensure that the desired effect
is
achieved. The need for stable, long-term dosing with GnRH agonists had led to
the
development of a small number of sustained release formulations, particularly
of the
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analogue Leuprolide (see above). The most recently introduced Leuprolide based
product is Eligard (Atrix Laboratories), which comprises a poly(DL-lactide-co-
glycolide) (PLG) polymer formulation dissolved in N -methyl-pyrrolidone (NMP),
to which leuprolide is added shortly before administration. Eligard is
available as 1-
month, 3-month and 4-month slow-release products. The principal disadvantages
of
this delivery system are in the nature of the delivery vehicle and the
complexity of
administration. A leuprolide depot product called Lupron and a triptorelin
depot
product called Trelstar LA are also available, which are administered as
suspensions
of PLGA microspheres. Again, the administration is complex and the nature of
the
depots is not ideal.
In particular, the Eligard system is supplied in two joinable syringes,
contained in a
kit including three pouches, stoppers, needles and replacement plunger-rods.
The
composition can be stored for only 5 days without refrigeration and must be
made
up and administered within 30 minutes. The mixing and administration requires
some 17 separate steps, including removing and replacing syringe plunger rods,
joining the two syringes and mixing by transferring the contents repeatedly
between
them. Evidently, this method requires a medical practitioner of considerable
experience for successful administration, and even in practiced hands will
take a
significant time to carry out the procedure. It would be a considerable
advantage,
therefore, to provide a depot product of similar performance in a "ready to
administer" form, preferably as a simple injectable liquid which could be
administered directly by no more than routine injection technique.
Existing GnRH analogue slow-release formulations must also be administered by
means of a sizable needle, typically of 20-gauge or wider. This is necessary
as a
result of the nature of the polymeric dosing systems used, which are typically
polymer dispersions or suspensions of polymeric microspheres. Evidently, it
would
be an advantage to provide a system of low viscosity, homogeneous solution,
which
could be administered easily through a narrow needle, thus decreasing the
discomfort of the patient during the procedure. Reducing preparation time of
health-
care professionals or patients prior to the actual administration to the
patient is
another important issue.
The poly-lactate, poly-glycolate and poly-lactate-co-glycolate polymers
typically
used for degrading slow-release formulations are also the cause of some
irritation in
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at least some patients. In particular, these polymers typically contain a
certain
proportion of acetic acid impurity, which will irritate the injection site on
administration. When the polymer then breaks down, lactic acid and glycolic
acid
are the degradation products so that further irritation is caused. As a result
of the
combined effects of wide-needle administration and irritant contents, the
discomfort
at the site of administration and the formation of connective scar tissue are
greater
than desirable.
A further limitation of the existing GnRH analogue depot systems is that
dosing
cannot easily be tailored to suit particular patients. One particular
indication for
which GnRH analogues have been shown as effective is in the delaying of
precocious puberty, but in this indication, there is a considerable variation
in subject
weight and a weight-tailored dose must be used. A depot system comprising a
pre-
weighted dry powder, which is dissolved or dispersed in an injection vehicle
by
means of a pair of linked syringes, does not allow any such control, however,
unless
a considerable range of pre-measured doses is to be provided. The depot
formulation cannot be partially administered because the dissolution of the
active
agent within the polymer solution may not be even. It would thus be a
considerable
advantage to have a homogeneous depot precursor, which allowed administration
of
a dose to be decided on a subject-specific basis at the time of
administration.
From a drug delivery point of view, polymer depot compositions generally have
the
disadvantage of accepting only relatively low drug loads and having a
"burst/lag"
release profile. The nature of the polymeric matrix, especially when applied
as a
solution or pre-polymer, causes an initial burst of drug release when the
composition
is first administered. This is followed by a period of low release, while the
degradation of the matrix begins, followed finally by an increase in the
release rate
to the desired sustained profile. This burst/lag release profile can cause the
in vivo
concentration of active agent to burst above the functional window immediately
following administration, then drop back through the bottom of the functional
window during the lag period before reaching a sustained functional
concentration.
Evidently, from a functional and toxicological point of view this burst/lag
release
profile is undesirable and could be dangerous. It may also limit the
equilibrium
concentration which can be provided due to the danger of adverse effects at
the
"peak" point.
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Evidently, in the case of GnRH agonists, the time of the "burst" period,
immediately
after administration is the time when the composition is having precisely the
opposite effect to that desired once equilibrium has established. When first
administered, the agonistic properties cause a transient increase in steroid
hormone
production, which, in the case of advanced prostate cancer, for example, can
cause
an exacerbation of the symptoms of urinary problems or even paralysis. There
have
been reports of patients who have died as a result of this initial phase, even
without
any "burst" effect and so evidently it would be preferable to avoid a burst
before
maintenance is established. Moreover, an unnecessary high quantity of peptide
is
given to patients in the initial "burst" phase possibly resulting in toxic
effects and an
increased cost of goods.
The manufacture of PLGA microbeads and suspensions is additionally a
considerable difficulty with certain existing depot systems. In particular,
since the
beads are particulate, and polymers clog membranes, they cannot generally be
sterile-filtered and furthermore, since the PLGA copolymer melts at around 40
C,
they cannot be heat-treated for sterility. As a result, a complex
manufacturing
process must all be conducted under conditions of high sterility.
The present inventors have now established that by providing a pre-formulation
comprising certain amphiphilic components, at least one GnRH analogue and a
biologically tolerable solvent in a low viscosity phase, such as molecular
solution, a
pre-formulation may be generated addressing many of the shortfalls of previous
GnRH analogue depot formulations. In particular, the pre-formulation is easy
to
manufacture, may be sterile-filtered, has low viscosity (allowing easy and
less
painful administration typically through a narrow needle), allows a high level
of
bioactive agent to be incorporated (thus potentially allowing a smaller amount
of
composition to be used), requires shallower injection and/or forms a desired
non-
lainellar depot composition in vivo having a controllable "burst" or "non-
burst"
release profile. The compositions are also formed from materials that are non-
toxic,
biotolerable and biodegradable, which can be administered by i.m., s.c. and
various
cavities.
In a first aspect, the present invention thus provides a pre-formulation
comprising a
low viscosity mixture of:
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a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) at least one GnRH analogue;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact witli an aqueous fluid.
Generally, the aqueous fluid will be a body fluid particularly extra-vascular
fluid,
extracellular fluid/interstitial fluid or plasma, and the pre-forinulation
will form a
liquid crystalline phase structure when contacted with such a fluid (e.g. in
vivo).
The pre-fonnulation of the invention will generally not contain any
significant
quantity of water prior to administration.
In a second aspect of the invention, there is also provided a method of
delivery of a
GnRH analogue to a human or non-human animal (preferably mammalian) body,
this method comprising parenterally administering (e.g. i.m. or preferably
s.c.) a pre-
formulation comprising a low viscosity mixture of
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) at least one GnR.H analogue;
whereby to form at least one liquid crystalline phase structure upon contact
with an
aqueous fluid in vivo following administration. Preferably, the pre-
formulation
administered in such a method is a pre-formulation of the invention as
described
herein.
In a further aspect, the present invention also provides a method for the
preparation
of a liquid crystalline depot composition comprising exposing a pre-
formulation
comprising a low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) at least one GnRH analogue;
to an aqueous fluid in vivo.
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Preferably the pre-formulation administered is a pre-formulation of the
present
invention as described herein.
In a still further aspect the present invention provides a process for the
formation of
a pre-formulation suitable for the administration of a bioactive agent to a
(preferably
mammalian) subject, said process comprising forming a low viscosity mixture of
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
and dissolving or di~persing at least one GnRH analogue in the low viscosity
mixture, or in at least one of coinponents a, b or c prior to forming the low
viscosity
mixture. Preferably the pre-formulation so-formed is a formulation of the
invention
as described herein.
In a yet still further aspect the present invention provides the use of a low
viscosity
mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) at least one GnRH analogue;
in the manufacture of a pre-formulation for use in the sustained
administration of
said GnRH analogue, wherein said pre-formulation is capable of forming at
least
one liquid crystalline phase structure upon contact with an aqueous fluid.
In a still further aspect, the present invention provides a method for the
treatment of
a human or non-human mammalian subject in need thereof with a GnRH analogue,
said method comprising administering to said subject a pre-formulation
comprising
a low-viscosity mixture of;
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) at least one GnRH analogue;
Preferably, the method of treatment is a method for the treatment of at least
one
condition selected from neoplastic diseases, including cancers, especially of
the
breast and prostate, and benign prostatic hypertrophy; premature or delayed
puberty
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in adolescents; hirsuitism; alzheimer's disease; and certain conditions
relating to the
reproductive system, such as hypogonadism, anovulation, amenorrhea,
oligospermia, endometriosis, leiomyomata (uterine fibroids), premenstral
syndrome,
and polycystic ovarian disease. The method may also be an in vitro
fertilisation
method (IVF).
In a yet further aspect, the present invention provides the use of;
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one qxygen containing organic solvent;
d) at least one GnRH analogue;
in the manufacture of a low viscosity pre-formulation medicament for use in
the in
vivo formation of a depot for treatment of neoplastic diseases, including
cancers,
especially of the breast and prostate, and benign prostatic hypertrophy;
premature or
delayed puberty in adolescents; hirsuitism; alzheimer's disease; and certain
conditions relating to the reproductive system, such as hypogonadism,
anovulation,
amenorrhea, oligospermia, endometriosis, leiomyomata (uterine fibroids),
premenstral syndrome, or polycystic ovarian disease, or for use as part of IVF
treatment.
The pre-formulations of the present invention are highly advantageous in that
they
are stable to prolonged storage in their final "administration ready" form. As
a
result, they may readily be supplied for administration either by healtli
professionals
or by patients or their carers, who need not be fully trained health
professionals and
may not have the experience or skills to make up complex preparations.
In a yet further aspect, the present invention provides a disposable
administration
device (which is also to include a device component) pre-loaded with a
measured
dose of a pre-formulation of the present invention. Such a device will
typically
contain a single dose ready for administration and will generally be sterile-
packed
such that the composition is stored within the device until administration.
Suitable
devices include cartridges, ampoules and particularly syringes and syringe
barrels,
either with integral needles or with standard (e.g. luer) fittings adapted to
take a
suitable disposable needle.
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The pre-filled devices of the invention may also suitably be included in an
administration kit, which kit also forms a further aspect of the invention. In
a still
further aspect, the invention thus provides a kit for the administration of at
least one
GnRH analogue, said kit containing a measured dose of a formulation of the
invention and optionally an administration device or component thereof.
Preferably
the dose will be held within the device or component, which will be suitable
for i.m.
or preferably s.c. administration. The kits may include additional
administration
components such as needles, swabs, etc. and will optionally and preferably
contain
instructions for administration. Such instructions will typically relate to
adininistration by a route as describe herein and/or for the treatment of a
disease
indicated herein above.
The formulations of the present invention generate a non-lamellar liquid
crystalline
phase following administration. The use of non-lamellar phase structures (such
as
liquid crystalline phases) in the delivery of bioactive agents is now
relatively well
established. Such structures form when an amphiphilic compound is exposed to a
solvent because the ainphiphile has both polar and apolar groups which cluster
to
form polar and apolar regions. These regions can effectively solubilise both
polar
and apolar compounds. In addition, many of the structures formed by
amphiphiles
in polar and/or apolar solvents have a very considerable area of polar/apolar
boundary at which other amphiphilic compounds can be adsorbed and stabilised.
Amphiphiles can also be formulated to protect active agents, to at least some
extent,
from aggressive biological environments, including enzymes, and thereby
provide
advantageous control over active agent stability and release.
The formation of non-lamellar regions in the amphiphile/water, amphiphile/oil
and
amphiphile/oil/water phase diagrams is a well known phenomenon. Such phases
include liquid crystalline phases such as the cubic P, cubic D, cubic G and
hexagonal phases, which are fluid at the molecular level but show significant
long-
range order, and the L3 phase which comprises a multiply interconnected bi-
continuous network of bilayer sheets which are non-lamellar but lack the long-
range
order of the liquid crystalline phases. Depending upon their curvature of the
amphiphile sheets, these phases may be described as normal (mean curvature
towards the apolar region) or reversed (mean curvature towards the polar
region).
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The non-lamellar liquid crystalline and L3 phases are thermodynamically stable
systems. That is to say, they are not simply a meta-stable state that will
separate
and/or reform into layers, lamellar phases or the like, but are the stable
thermodynamic form of the lipid/solvent mixture.
It is important that the pre-formulations of the invention are not liquid
crystalline
prior to administration because bulk liquid crystalline phase is generally
highly
viscous. The pre-formulations are thus low viscosity, non-liquid-crystalline
formulations which undergo a phase change upon adininistration to form a
liquid
crystalline mass. Particularly preferred examples of low viscosity mixtures
are
molecular solutions and/or isotropic phases such as L2 and/or L3 phases. As
describe above, the L3 is a non-lamellar phase of interconnected sheets which
has
some phase structure but lacks the long-range order of a liquid crystalline
phase.
Unlike liquid crystalline phases, which are generally highly viscous, L3
phases are
of lower viscosity. Obviously, mixtures of L3 phase and molecular solution
and/or
particles of L3 phase suspended in a bulk molecular solution of one or more
components are also suitable. The L2 phase is the so-called "reversed
micellar"
phase or microemulsion. Most preferred low viscosity mixtures are molecular
solutions, L3 phases and mixtures thereof. L2 phases are less preferred,
except in
the case of swollen L2 phases as described below.
As used herein, the term "low viscosity mixture" is used to indicate a mixture
which
may be readily administered to a subject and in particular readily
administered by
means of a standard syringe and needle arrangement. This may be indicated, for
example by the ability to be dispensed from a 1 ml disposable syringe through
a
small gauge needle. Preferably, the low viscosity mixtures can be dispensed
through
a needle of 19 awg, preferably smaller than 19 gauge, more preferably 23 awg
(or
most preferably even 27 gauge) needle by manual pressure. In a particularly
preferred embodiment, the low viscosity mixture should be a mixture capable of
passing through a standard sterile filtration membrane such as a 0.22 m
syringe
filter. A typical range of suitable viscosities would be, for example, 0.1 to
5000
mPas, preferably 1 to 1000 mPas at 20 C.
It has been observed that by the addition of small amounts of low viscosity
solvent,
as indicated herein, a very significant change in viscosity can be provided.
As
indicated in Figure 1, for example, the addition of only 5% solvent to a lipid
mixture
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can reduce viscosity 100-fold and addition of 10% may reduce the viscosity up
to
10,000 fold. In order to achieve this non-linear, synergistic effect, in
lowering
viscosity it is important that a solvent of appropriately low viscosity and
suitable
polarity be employed. Such solvents include those described herein infra.
The present invention provides a pre-formulation comprising components a, b, c
and
at least one GnRH analogue as indicated herein. The amounts of these
components
will typically be in the range 40-70% a), 30-60% b) and 0.1-10% c), with the
GnRH
analogue present at 0.1 % to 10%. All % being by weight herein throughout,
unless
otherwise indicated., The formulations may consist of essentially only these
components and in one aspect consist entirely of such coinponents. Preferable
ranges for component a) are 43-60%, particularly 45-55 and preferable ranges
of
component b) are 35-55%, particularly 40 to 50%.
Ratios of a:b are typically 40:60 to 70:30, preferably 45:55 to 60:40 and more
preferably 48:52 to 55:45. Ratios of around 50:50 are highly effective.
The amount of solvent component c) in the preformulation will have a
considerable
effect upon several features. In particular, the viscosity and the rate (and
duration)
of release will alter significantly with the solvent level. The amount of
solvent will
thus be at least sufficient to provide a low viscosity mixture but will
additionally be
determined so as to provide the desired release rate. This may be determined
by
routine methods in view of the Exainples below. Typically a level of 0.1 to
10%
solvent will provide suitable release and viscosity properties. This will
preferably
be 2 to 8% and an amount of around 5% is highly effective.
It is the remarkable finding of the present inventors that the proportion of
solvent in
the formulation can be used to "tune" the release profile of the active agent
during
the first few days of release. In particular, although all formulations of the
invention
have a surprisingly low "burst/lag" effect (in fact there are may be no lag
period at
all), and reach a plateau release level within a few days (e.g. 5 days,
preferably 3
days, more preferably 1 day) of injection, if a controlled "burst"/initial
release of
active agent is required in the first 1-2 days then this can be provided by
increasing
the solvent proportion to the upper region of the range given above. In
contrast, in
the mid- to lower-region of the range, a formulation giving a depot with
essentially
no burst and a rapid decline to the plateau release level is provided.
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Thus, in one embodiment, the present invention provides formulations and
depots
containing around 0.1 to 6 wt% component c) and having a low release of the
active
compound during the first days after administration ("non-burst profile"). In
an
alternative embodiment, the present invention provides formulations and depots
containing around 6.5 to 10 wt% component c) and having high initial release
of the
active compound during the first days after administration ("burst profile").
The low initial release ("non-burst profile") of active agent is defined such
that the
area under a plasma, concentration against time the curve during the first 24
hours is
less than 15% of the area under the curve for the entire curve (measured or
extrapolated from time 0 to infinity or from time 0 to the last sampling time
point),
more preferably less than 10% and most preferable less than 7%. In addition,
the
decline to plateau plasma concentration levels after the initial peak should
be rapid,
such that plateau is reached with in 48 hours, more preferably within 24
hours, and
most preferably witllin 12 hours. Conversely, a high initial release ("burst
profile")
is such that more than 15% of active agent is released within 24 hours and
more
preferably more than 20 % is released during the first 24 hours. The decline
to
plateau will not occur until after 36 hours, more preferably after 48 hours
and most
preferably after 72 hours. It is preferable that each of these profiles is
combined
with a rapid settling of the plasma active agent concentration to "plateau"
level. For
example, the plasma concentration after 10 days should be no more than 50%
greater or less than the average concentration over days 5 to 20. Preferably
this will
be no more than 30% and more preferably no more than 20%.
As indicated above, the amount of component c in the pre-formulations of the
invention will be at least sufficient to provide a low viscosity mixture (e.g.
a
molecular solution, see above) of components a, b and c and will be easily
determined for any particular combination of components by standard methods.
The
phase behaviour itself may be analysed by techniques such as visual
observation in
combination with polarized light microscopy, nuclear magnetic resonance, and
cryo-
transmission electron microscopy (cryo-TEM) to look for solutions, L2 or L3
phases, or liquid crystalline phases or as in the case of cryoTEM, dispersed
fragments of such phases. Viscosity may be measured directly by standard
means.
As described above, an appropriate practical viscosity is that which can
effectively
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be syringed and particularly sterile filtered. This will be assessed easily as
indicated
herein.
Component "a" as indicated herein is at least one diacyl glycerol (DAG) and
thus
has two non-polar "tail" groups. The two non-polar groups may have the same or
a
differing number of carbon atoms and may each independently be saturated or
unsaturated. Examples of non-polar groups include C6-C32 alkyl and alkenyl
groups,
which are typically present as the esters of long chain carboxylic acids.
These are
often described by reference to the number of carbon atoms and the number of
unsaturations in the,carbon chain. Thus, CX:Z indicates a hydrocarbon chain
having
X carbon atoms and Z unsaturations. Examples particularly include caproyl
(C6:0),
capryloyl (C8:0), capryl (C10:0), lauroyl (C12:0), myristoyl (C14:0),
palmitoyl
(C 16: 0), phytanoyl (C 16:0), palmitoleoyl (C 16:1), stearoyl (C 18 :0),
oleoyl (C 18 :1),
elaidoyl (C18:1), linoleoyl (C18:2), linolenoyl (C18:3), arachidonoyl (C20:4),
behenoyl (C22:0) and lignoceroyl (C24:9) groups. Thus, typical non-polar
chains
are based on the fatty acids of natural ester lipids, including caproic,
caprylic, capric,
lauric; myristic, palmitic, phytanic, palmitolic, stearic, oleic, elaidic,
linoleic,
linolenic, arachidonic, behenic or lignoceric acids, or the corresponding
alcohols.
Preferable non-polar chains are palmitic, stearic, oleic and linoleic acids,
particularly
oleic acid.
Mixtures of any number of diacyl-lipids may be used as component a. Preferably
this component will include at least a portion of glycerol dioleate (GDO). A
highly
preferred example is DAG comprising at least 50%, preferably at least 80% and
even comprising substantially 100% GDO.
Since GDO and other diacyl glycerols are products derived from natural
sources,
there is generally a certain proportion of "contaminant" lipid having other
chain
lengths etc. In one aspect, GDO as used herein is thus used to indicate any
commercial grade of GDO with concomitant impurities (i.e. GDO of commercial
purity). These impurities may be separated and removed by purification but
providing the grade is consistent this is rarely necessary. If necessary,
however,
"GDO" may be essentially chemically pure GDO, such as at least 80% pure,
preferably at least 85% pure and more preferably at least 90% pure GDO.
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Component "b" in the present invention is at least one phosphatidyl choline
(PC).
As with component a, this component comprises a polar head group and at least
one
non-polar tail group. The difference between components a and b lies
principally in
the polar group. The non-polar portions may thus suitably be derived from the
fatty
acids or corresponding alcohols considered above for component a. As with
component a), the PC will contain two non-polar groups.
The phosphatidyl choline portion, even more suitably than any diacyl glycerol
portion, may be derived from a natural source. Suitable sources of
phospholipids
include egg, heart (e.g. bovine), brain, liver (e.g. bovine) and plant sources
including
soybean. Such sources may provide one or more constituents of component b,
which may comprise any mixture of phospholipids. Any single PC or mixture of
PCs from these or other sources may be used, but mixtures comprising soy PC or
egg PC are highly suitable. The PC component preferably contains at least 50%
soy
PC or egg PC, more preferably at least 75% soy PC or egg PC and most
preferably
essentially pure soy PC or egg PC.
Since the pre-formulations of the invention are to be administered to a
subject for
the controlled release of a GnRH analogue active agent, it is important that
the
coinponents are biocompatible. In this regard, the pre-formulations of the
present
invention are highly advantageous since both PC and DAGs are well tolerated
and
are broken down in vivo into components that are naturally present in the
mammalian body.
A particularly favoured combination of components a and b are GDO with PC,
especially GDO with soy PC.
Component "c" of the pre-formulations of the invention is an oxygen containing
organic solvent. Since the pre-formulation is to generate a depot composition
following administration (e.g. in vivo), upon contact with an aqueous fluid,
it is
desirable that this solvent be tolerable to the subject and be capable of
mixing with
the aqueous fluid, and/or diffusing or dissolving out of the pre-formulation
into the
aqueous fluid. Solvents having at least moderate water solubility are thus
preferred.
In a preferred version, the solvent is such that a relatively small addition
to the
composition comprising a and b, i.e. preferably below 10%, give a large
viscosity
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reductions of one order of magnitude or more. As described herein, the
addition of
10% solvent can give a reduction of two, three or even four orders of
magnitude in
viscosity over the solvent-free composition, even if that composition is a
solution or
L2 phase containing no solvent, or an unsuitable solvent such as water, or
glycerol.
Typical solvents suitable for use as component c include at least one solvent
selected
from alcohols, ketones, esters (including lactones), ethers, amides and
sulphoxides.
Alcohols are particularly suitable and form the preferred class of solvents.
Examples of suitable alcohols include ethanol, isopropanol and glycerol
formal.
Ethanol is most preferred. Monools are preferred to diols and polyols. Where
diols
or polyols are used, this is preferably in combination with an at least equal
amount
of monool or other preferred solvent. Examples of ketones include acetone, n-
methyl pyrrolidone (NMP), 2-pyrrolidone, and propylene carbonate. Suitable
ethers
include diethylether, glycofurol, diethylene glycol monoethyl ether,
dimethylisobarbide, and polyethylene glycols. Suitable esters include ethyl
acetate
and isopropyl acetate and dimethyl sulphide is as suitable sulpliide solvent.
Suitable
amides and sulphoxides include dimethylacetamide (DMA) and dimethylsulphoxide
(DMSO), respectively.
A highly preferred combination is soy PC, GDO and ethanol.
It is preferable that little or none of component c contains halogen
substituted
hydrocarbons since these tend to have lower biocompatibility. Where a portion
of
halogenated solvent such as dichloromethane or chloroform is necessary, this
proportion will generally be minimised.
Component c as used herein may be a single solvent or a mixture of suitable
solvents but will generally be of low viscosity. This is important because one
of the
key aspects of the present invention is that it provides pre-formulations that
are of
low viscosity and a primary role of a suitable solvent is to reduce this
viscosity.
This reduction will be a combination of the effect of the lower viscosity of
the
solvent and the effect of the molecular interactions between solvent and lipid
composition. One observation of the present inventors is that the oxygen-
containing
solvents of low viscosity described herein have highly advantageous and
unexpected
molecular interactions with the lipid parts of the composition, thereby
providing a
non-linear reduction in viscosity with the addition of a small volume of
solvent.
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The viscosity of the "low viscosity" solvent component c (single solvent or
mixture)
should typically be no more than 18 mPas at 20 C. This is preferably no more
than
15 mPas, nlore preferably no more than 10 mPas and most preferably no more
than
7 mPas at 20 C.
A further advantage of the present pre-formulations is that a higher level of
bioactive agent may be incorporated into the system. In particular, by
appropriate
choice of components a-c (especially c), high levels of active agent may be
dissolved or suspended in the pre-formulations. This allows a reduction in the
administered volume and thus less discomfort to subjects.
The pre-formulations of the present invention typically do not contain
significant
amounts of water. Since it is essentially impossible to remove every trace of
water
from a lipid composition, this is to be taken as indicating that only such
minimal
trace of water exists as cannot readily be removed. Such an amount will
generally
be less than 1% by weight, preferably less that 0.5% by the weight of the pre-
formulation. In one preferred aspect, the pre-formulations of the invention do
not
contain glycerol, ethylene glycol or propylene glycol and contain no more than
a
trace of water, as just described.
The pre-formulations of the present invention contain one or more GnRH
analogues
or other active (see below)(which are intended by any reference to "active
agents"
herein). Since GnRH is a peptide hormone, typical GnRH analogues will be
peptides, especially of 12 or fewer amino acids. Preferably such peptides will
be
structurally related to GnRH I, II and/or III, and/or one or more of the known
analogues, including those listed here. Peptides may contain only amino acids
selected from those 20 a-aniino acids indicated in the genetic code, or more
preferably may contain their isomers and other natural and non-natural amino
acids,
(generally a, (3 or y amino acids) and their analogues and derivatives.
Preferred
amino acids include those listed above as constituents of the known GnRH
analogues.
Amino acid derivatives are especially useful at the termini of the peptides,
where the
terminal amino or carboxylate group may be substituted by or with any other
functional group such as hydroxy, alkoxy, carboxy, ester, amide, thio, amido,
amino,
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alkyl amino, di- or tri-alkyl amino, alkyl (by which is meant, herein
throughout C1-
C12 alkyl, preferably CI-C6 alkyl e.g. methyl, ethyl, n-propyl, isopropyl, n-
butyl, iso-
, sec- or t-butyl etc.), aryl (e.g phenyl, benzyl, napthyl etc) or other
functional
groups, preferably with at least one heteroatom and preferably having no more
than
10 atoms in total, more preferably no more than 6.
Particularly preferred GnRH analogues are constrained peptides of 6 to 12
alpha-
amino acids, of which particular examples include those indicated above, and
particularly leuprolide and goserelin, of the sequences indicated above.
By "GnRH analogues", as used herein is indicated any GnRH agonist or
antagonist,
preferably peptides, peptide derivatives or peptide analogues. Peptide derived
GnRH agonists are most preferred, such as those indicated above and especially
leuprolide or goserelin.
The GnRH analogue will generally be formulated as 0.02 to 12% by weight of the
total formulation. Typical values will be 0.1 to 10%, preferably 0.2 to 8% and
more
preferably 0.5 to 6%. A GnRH analogue content of around 1-5% is most
preferable.
Doses of the GnRH analogue suitable for inclusion in the formulation, and thus
the
volume of formulation used will depend upon the release rate (as controlled,
for
example by the solvent type and amount use) and release duration, as well as
the
desired therapeutic level, the activity of the specific agent, and the rate of
clearance
of the particular active chosen. Typically an amount of 0.1 to 500 mg per dose
would be suitable for providing a therapeutic level for between 7 and 180
days. This
will preferably be 1 to 200 mg. For leuprolide or goserelin, the level will
typically
be around 1 to 120 mg (e.g. for a 30 to 180 day duration). Preferably, the
amount of
leuprolide will be around 0.02 to 1 mg per day between injections, for depots
designed for release over 30 days to 1 year, preferably 3 to 6 months.
Evidently, the
stability of the active and linearity of the release rate will mean that the
loading to
duration may not be a linear relationship. A depot administered every 30 days
might
have, for example 2 to 30 mg or a 90 day depot have 6 to 90 mg of active, such
as
one of the GnRH analogues indicated herein.
The pre-formulations of the present invention are formulated to be
administered
parenterally. This administration will generally not be an intra-vascular
method but
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will preferably be subcutaneous, intracavitary or intramuscular. Typically the
administration will be by injection, which term is used herein to indicate any
method
in which the formulation is passed through the skin, such as by needle,
catheter or
needle-less injector.
Preferred parenteral administration is by i.m or s.c. injection, most
preferably by
deep s.c. injection. An important feature of the composition of the invention
is that it
can be administered both by i.m. and s.c. and otlier routes without toxicity
or
significant local effects. It is also suitable for intracavital
administration. The deep
s.c. injection has the advantage of being less deep and less painful to the
subject than
the (deep) i.m. injection used for some current depots and is technically most
suitable in the present case as it combines ease of injection with low risk of
local
side effects. It is a surprising observation of the present inventors that the
formulations provide sustained release of active agent over a predictable time
period
by both subcutaneous and intramuscular injection. This therefore allows the
site of
injection to be varied widely and allows the dose to be administered without
detailed
consideration of the tissue depth at the site of injection.
The pre-formulations of the present invention provide non-lamellar liquid
crystalline
depot compositions upon exposure to aqueous fluids, especially in vivo. As
used
herein, the term "non-lamellar" is used to indicate a normal or reversed
liquid
crystalline phase (such as a cubic or hexagonal phase) or the L3 phase or any
combination thereof. The term liquid crystalline indicates all hexagonal, all
cubic
liquid crystalline phases and/or all mixtures thereof. Hexagonal as used
herein
indicates "normal" or "reversed" hexagonal (preferably reversed) and "cubic"
indicates any cubic liquid crystalline phase unless specified otherwise.
For many combinations of lipids, only certain non-lamellar phases exist, or
exist in
any stable state. It is a surprising feature of the present invention that
compositions
as described herein frequently exhibit non-lamellar phases which are not
present
with many other combinations of components. In one particularly advantageous
embodiment, therefore, the present invention relates to compositions having a
combination of components for which an I2 and/or L2 phase region exists when
diluted with aqueous solvent. The presence or absence of such regions can be
tested
easily for any particular combination by siniple dilution of the composition
with
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aqueous solvent and study of the resulting phase structures by the methods
described
herein.
In a highly advantageous embodiment, the compositions of the invention may
form
an I2 phase, or a mixed phase including I2 phase upon contact witli water. The
I2
phase is a reversed cubic liquid crystalline phase having discontinuous
aqueous
regions. This phase is of particular advantage in the controlled release of
active
agents and especially in combination with polar active agents, such as water
soluble
actives because the discontinuous polar domains prevent rapid diffusion of the
actives. Depot precursors in the L2 are highly effective in combination with
an 12
phase depot formation. This is because the L2 phase is a so-called "reversed
micellar" phase having a continuous hydrophobic region surrounding discrete
polar
cores. L2 thus has similar advantages with hydrophilic actives.
In transient stages after contact with body fluid the composition can comprise
multiple phases since the formation of an initial surface phase will retard
the passage
of solvent into the core of the depot, especially with substantial sized
admiriistrations of internal depots. Without being bound by theory, it is
believed
that this transient formation of a surface phase, especially a liquid
crystalline surface
phase, serves to dramatically reduce the "burst/lag" profile of the present
compositions by immediately restricting the rate of exchange between the
composition and the surroundings. Transient phases may include (generally in
order
from the outside towards the centre of the depot): Hii or La,, IZ, L2, and
liquid
(solution). It is highly preferred that the composition of the invention is
capable
forming at least two and more preferably at least three of these phases
simultaneously at transient stages after contact with water at physiological
temperatures. In particular, it is highly preferred that one of the phases
formed, at
least transiently, is the 12 phase.
It is important to appreciate that the pre-formulations of the present
invention are of
low viscosity. As a result, these pre-formulations must not be in any bulk
liquid
crystalline phase since all liquid crystalline phases have a viscosity
significantly
higher than could be administered by syringe or spray dispenser. The pre-
forinulations of the present invention will thus be in a non-liquid
crystalline state,
such as a solution, L2 or L3 phase, particularly solution or L2. The L2 phase
as used
herein throughout is preferably a "swollen" L2 phase containing greater than
10 wt%
of solvent (component c) having a viscosity reducing effect. This is in
contrast to a
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"concentrated" or "unswollen" L2 phase containing no solvent, or a lesser
amount of
solvent, or containing a solvent (or mixture) which does not provide the
decrease in
viscosity associated with the oxygen-containing, low viscosity solvents
specified
herein.
Upon administration, the pre-formulations of the present invention undergo a
phase
structure transition from a low viscosity mixture to a high viscosity
(generally tissue
adherent) depot composition. Generally this will be a transition from a
molecular
mixture, swollen L2 and/or L3 phase to one or more (high viscosity) liquid
crystalline phases such as normal or reversed hexagonal or cubic liquid
crystalline
phases or mixtures thereof. As indicated above, further phase transitions may
also
take place following administration. Obviously, complete phase transition is
not
necessary for the functioning of the invention but at least a surface layer of
the
administered mixture will form a liquid crystalline structure. Generally this
transition will be rapid for at least the surface region of the administered
formulation
(that part in direct contact with air, body surfaces and/or body fluids). This
will
most preferably be over a few seconds or minutes (e.g. up to 30 minutes,
preferably
up to 10 minutes, more preferably 5 minutes of less). The remainder of the
composition may change phase to a liquid crystalline phase more slowly by
diffusion and/or as the surface region disperses.
In one preferred embodiment, the-present invention thus provides a pre-
formulation
as described herein of which at least a portion forms a hexagonal liquid
crystalline
phase upon contact with an aqueous fluid. The thus-formed hexagonal phase may
gradually disperse and/or degrade, releasing the active agent, or may
subsequently
convert to a cubic liquid crystalline phase, which in turn then gradually
disperses. It
is believed that the hexagonal phase will provide a more rapid release of
active
agent, in particular of hydrophilic active agent, than the cubic phase
structure,
especially the IZ and L2 phase. Thus, where the hexagonal phase forms prior to
the
cubic phase, this will result in an initial release of active agent to bring
the
concentration up to an effective level rapidly, followed by the gradual
release of a
"maintenance dose" as the cubic phase degrades. In this way, the release
profile
may be controlled.
Without being bound by theory, it is believed that upon exposure (e.g. to body
fluids), the pre-formulations of the invention lose some or all of the organic
solvent
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included therein (e.g. by diffusion) and take in aqueous fluid from the bodily
environment (e.g. the in vivo environment) such that at least a part of the
formulation generates a non-lamellar, particularly liquid crystalline phase
structure.
In most cases these non-laniellar structures are highly viscous and are not
easily
dissolved or dispersed into the in vivo environment. The result is a
monolithic
"depot" generated in vivo with only a limited area of exposure to body fluids.
Furthermore, because the non-lamellar structure has large polar, apolar and
boundary regions, it is highly effective in solubilising and stabilising
active agents
such as peptides and protecting these from degradation mechanisms. As the
depot
composition formed from the pre-formulation gradually degrades over a period
of
days, weeks or months, the active agent is gradually released and/or diffuses
out
from the composition. Since the environment within the depot composition is
relatively protected, the pre-formulations of the invention are highly
suitable for
active agents with a relatively low biological half-life (see above).
The depot systems formed by the formulations of the present invention are
highly
effective in protecting the active agent from degradation and thus allow an
extended
release period. Comparative tests have been carried out between the known PLGA
slow-release product and formulations of the present invention containing GDO,
soy
PC, ethanol and active agents. These indicate that formulations of the present
invention give lesser degradation under simulated in vivo conditions than
known
compositions. The formulations of the invention thus may provide in vivo
depots of
GnRH analogues which require administration only once every 20 to 360 days,
preferably 30 to 240 days, more preferably 60 to 180 days. Evidently, a longer
stable release period is desirable for patient comfort and compliance, as well
as
demanding less time from health professionals.
A considerable advantage of the depot precursors of the present invention is
that
they are stable homogeneous phases. That is to say, they may be stored for
considerable periods (preferably at least 6 months) at room or refrigerator
temperature, without phase separation. As well as providing advantageous
storage
and facile administration, this allows for the dose of GnRH analogue to be
selected
by reference to the species, age, sex, weight, and/or physical condition of
the
individual subject, by means of injecting a selected volume. Furthermore, the
present inventors have surprisingly found that the initial release of active
agent
(observed as Cma,) is not proportional to dose volume, in ranges of at least
10-fold in
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sample volume injection, while the total drug exposure (observed as AUC or
mean
plateau plasma concentration) is proportional to the injection volume. On the
contrary, it has been shown that Cn,,,x can be correlated to the surface area
of the
injected dose volume. That is, C,,,a, is proportional to the two-third power
of the
injected dose volume. Increasing the dose volume by a factor of 10 will not
increase
the C,aX 10 times and the relationship between C,,,a, and the total drug
exposure
(AUC or mean plateau plasma concentration level) will thus decrease with
increasing dose volume. This is highly advantageous, because this property
reduce
the risk of reaching potentially toxic plasma drug concentrations even if the
total
dose is significantly, increased. Even in situations where dosing is not
directly
proportional to injection volume, however, the homogenous nature of the depot
precursors importantly allow for partial administration of a pre-measured dose
and
this administration may be made by reference to a dosing table, chart,
software
calculation etc. which may take into account any or all relevant subject
variables.
The present invention thus provides for methods comprising the selection of a
dosing amount specific to an individual, particularly by subject weight. The
means
for this dose selection being by administration voluine.
Furthermore, the PLGA solution-type depots formulated for long-term (e.g. 4-
month) release of leuprolide show a"burst" release of the active, with a
maximum
concentration during the initial burst being 100-600 times that of the plateau
level.
As indicated above, this can have considerably disadvantages, especially with
GnRH
agonists, since their initial effect is to increase gonadal steroid
production, which
can cause symptoms to worsen.
It is an unexpected finding of the present inventors that the pre-formulations
result
in a depot composition that have very little "burst" effect in the active
agent release
profile. This is unexpected because it might be expected that the low
viscosity
mixture (especially if this is a solution) of the pre-composition would
rapidly lose
active agent upon exposure to water in the way that is observed for PLGA
suspended in NMP. In fact, pre-formulations of the invention have shown
considerably less of an initial "burst" than previously known polymer-base
depot
compositions which tend to have an initial "wash off' or "wash out"of surface-
bound or dissolved active agent. In one embodiment, the invention thus
provides
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injectable pre-formulations and resulting depot compositions wherein the
highest
plasma concentration of active after administration is no more than 40 times
the
average concentration between 24 hours and 5 days of administration. This
ratio is
preferably no more than 25 times and most preferably no more than 20 times
(e.g.
up to 10 or up to 5 times) the average concentration. This is an improvement
of an
order of magnitude over the existing PLGA/NMP depot product.
The compositions of the invention also allow for the generation of depot
compositions with very little "lag" effect after administration. In a further
embodiment, the invention thus provides injectable pre-formulations and
resulting
depot compositions wherein the plasma concentration of active at 7 days after
a
single administration is no lower than the plasma concentration of active at
21 days
after administration. Similarly, the concentration of active should be higher
at all
times in the first 21 days than the concentration at any time from 30 days
after
administration onwards. This gradually decaying release profile has not
previously
been demonstrated for slow release GnRH analogue formulation.
In combination witli the features and preferred features indicated herein, the
pre-
formulations of the invention may have one or more of the following preferred
features independently or in combination:
Component a) comprises, consists essentially of or preferably consists of GDO;
Component b) comprises, consists essentially of or preferably consists of soy
PC;
Component c) comprises, consists essentially of or preferably consists of a 1,
2, 3 or
4 carbon alcohol, preferably isopropanol or more preferably ethanol;
The pre-formulation contains at least one GnRH analogue selected from those
indicated herein, preferably leuprolide, or goserelin;
The pre-formulation has a low viscosity as indicated herein.
The pre-formulation forms a liquid crystalline phase as indicated herein upon
in vivo
administration.
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The pre-formulation generates a depot following in vivo administration, which
depot
releases at least one GnRH analogue at a therapeutic level over a period of at
least
30 days, preferably at least 90 days, more preferably at least 180 days.
In combination with the features and preferred features indicated herein, the
method(s) of treatment of the present invention may have one or more of the
following preferred features independently or in combination:
The method comprises the administration of at least one formulation with one
or
more preferred feaWres as indicated above;
The method con7prises the administration of at least one formulation as
indicated
herein by i.m., s.c. or preferably deep s.c. injection;
The method comprises administration by means of a pre-filled administration
device
as indicated herein;
The method comprises administration through a needle no larger than 20 gauge,
preferably smaller than 20 gauge, and most preferably 23 gauge or smaller;
The method comprises a single administration every 20 to 360 days, preferably
30 to
240 days, more preferably 60 to 180 days.
In combination with the features and preferred features indicated herein, the
use(s)
of the pre-formulations indicated herein in the manufacture of inedicaments
may
have one or more of the following preferred features independently or in
combination:
The use comprises the use of at least one formulation with one or more
preferred
features as indicated above;
The use comprises the manufacture of a medicament for administration of at
least
one formulation as indicated herein by i.m., s.c. or preferably deep s.c.
injection;
The use comprises the manufacture of a medicament for administration by means
of
a pre-filled administration device as indicated herein;
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The use comprises the manufacture of a medicament for administration through a
needle no larger than 20 gauge, preferably smaller than 20 gauge, and most
preferably 23 gauge or smaller;
The use comprises the manufacture of a medicament for administration once
every
20 to 360 days, preferably 30 to 240 days, more preferably 60 to 180 days.
In combination with the features and preferred features indicated herein, the
pre
filled devices of the,invention may have one or more of the following
preferred
features independently or in combination:
They contain a preferred formulation as indicated herein;
They comprise a needle smaller than 20 gauge, preferably no larger than 23
gauge;
They contain a single dose of 0.1 to 500 mg of GnRH analogue, preferably 1 to
200
mg;
They contain goserelin or leuprolide, at around 5 to 90 mg ;
They contain a homogeneous mixture of a composition of the invention in ready-
to-
inject form.
They contain a total volume for administration of no more than 5 ml,
preferably no
more than 3 ml more preferably no more than 2 ml.
In combination with the features and preferred features indicated herein, the
kits of
the invention may have one or more of the following preferred features
independently or in combination:
They contain a preferred formulation as indicated herein;
They contain a pre-filled device as indicated herein;
They contain a needle smaller than 20 gauge, preferably no larger than 23
gauge;
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They contain a single dose of 0.1 to 500 mg of GnRH analogue, preferably 1 to
200
mg;
They contain leuprolide or goserelin, at around 5 to 90 mg ;
They contain a total volume for administration of no more than 5 ml,
preferably no
more than 3 ml more preferably no more than 2 ml.
They contain instructions for administration by a route and/or at a frequency
as
indicated herein;
They contain instructions for administration for use in a method of treatment
as
described herein.
In Further aspects of the present invention, corresponding depot pre-
formulations
may be made using alternative active agents. For each of these, the types and
proportions of components a), b) and c) will be as indicated above as general
and
preferred formulations, particularly as indicated in the attached claims, in
which the
appropriate doses of active agents indicated below may be substitued in place
of the
GnRH analogues. The formulations may be generated, tested and used by methods
analogous to those for the GnRH analogues considered above, as is demonstrated
in
the attached examples. All aspects of the invention relating to compositions,
kits
and devices apply equally to the following active agents and methods of
treatment
apply as indicated below.
In one aspect, the invention thus provides a pre-formulation, comprising a low
viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) risperidone or at least one analogue or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
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The risperidone analogue composition will preferably be a preferred
composition as
indicated herein. The content of risperidone or at least one analogue or
derivative
thereof will typically be around 1 to 200 mg per week of depot duration,
preferably
to 100 mg per week duration for a duration of 1 to 12 weeks.
5
The invention further provides a method of medical treatement comprising
administration of a risperidone analogue composition as described above,
preferably
a method for the treatment of schizophrenia. The invention also provides the
use of
a risperidone analogue composition as described above in the manufacture of a
10 medicament for the ,treatment of schizophrenia. The invention additionally
provides
for a pre-filled administration device as indicated herein and a kit as
indicated herein
comprising the risperidone analogue composition.
An additional and surprsing aspect of the compositions of the present
invention is
that they can from depots for the release of non-polar small (i.e. non-
peptide)
molecules over a sustained period of at least 7 days, preferably at least 2
weeks and
more preferably at least 4 weeks. Previously known depot compositions have
generally been limited to peptides and other polar active agents and agents of
high
molecular weight. This applies particularly to lipid based depot compositions
which
have not shown sustained release of non-polar and.or small molecule actives
for
more than a few days previously. In this aspect of the invention, by "small
molecule" is meant an active agent of niolecular weight below 1,000 amu,
preferably
less than 800 and most preferably less than 500 amu. by non -polar is
indicated a
molecule with a "logP" partition coefficient between octanol and water of
greater
than 1, preferably greater than 2 and more preferably greater than 3. Steroid
hormones such as testosterone are particularly favoured non-polar small
molecules.
The compositional region of around 60:40 to 40:60 a:b has been found to be the
most appropriate for sustained release, for all of the actives mentioned
herein, but
particularly for these types of active agents. This is preferably the region
55:45 to
45:55, most preferably 52:48 to 48:52 a:b (by
weight).
In a further aspect, the invention thus provides a pre-formulation, comprising
a low
viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
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c) at least one oxygen containing organic solvent;
d) testosterone or at least one analogue or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
The testosterone analogue composition will preferably be a preferred
composition as
indicated herein. The content of testosterone or at least one analogue or
derivative
thereof will typically be around 5 to 100 mg per week of depot duration,
preferably
to 70 mg per week duration for a duration of 1 to 24, preferably 8 to 16 (e.g.
12)
10 weeks.
The invention further provides a method of treatment comprising administration
of a
testosterone analogue coinposition as described above, especially in a subject
in
need thereof. The method of treatment is particularly for the treatment of
male
hypogonadism. The invention also provides the use of a testosterone analogue
composition as described above in the manufacture of a medicament for the
treatment of male hypogonadism. The invention additionally provides for a pre-
filled administration device as indicated herein and a kit as indicated herein
comprising the testosterone analogue composition.
In another aspect, the invention thus provides a pre-formulation, comprising a
low
viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) at least one aromatase inhibitor;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
The aromatase inhibitor will typically be anastrazole, femara, or aromasin.
The
aromatase inhibitor composition will preferably be a preferred composition as
indicated herein. The content of at least one aromatase inhibitor will
typically be
equivalent to a 3 to 10 mg oral dose per week of depot duration, which will
typically
be 0.03 to 1 mg pre week, preferably 0.05 to 0.8 mg per week duration, for a
duration of 1 to 24, preferably 4 to 12 (e.g. 8) weeks.
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The invention further provides a method of treatment comprising administration
of
an aromatase inhibitor composition as described above, especially in a subject
in
need thereof. The method of treatment is particularly for the treatment of
early,
locally advanced or metastatic breast cancer. The invention also provides the
use of
an aromatase inhibitor composition as described above in the manufacture of a
medicament for the treatment of early, locally advanced or metastatic breast
cancer.
The invention additionally provides for a pre-filled administration device as
indicated herein and a kit as indicated herein comprising the aromatase
inhibitor
composition.
In a further aspect, the invention also provides a pre-formulation, comprising
a low
viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) buprenorphine or at least one analogue or derivative thereof;
wlierein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
The buprenorphine analogue composition will preferably be a preferred
composition
as indicated herein. The content of buprenorphine or at least one analogue or
derivative thereof will typically be around 10 to 250 mg per week of depot
duration,
preferably 15 to 200 mg per week duration for a duration of 1 to 24,
preferably 4 to
12 (e.g. 8) weeks.
The invention further provides a method of treatment comprising administration
of a
buprenorphine analogue composition as described above, especially in a subject
in
need thereof. The method of treatment is particularly for the treatment of
pain,
especially chronic pain, or in the treatment of opioid addiction. The
invention also
provides the use of a buprenorphine analogue composition as described above in
the
manufacture of a medicament for the treatment of for the treatment of pain,
especially chronic pain, or in the treatment of opioid addiction. The
invention
additionally provides for a pre-filled administration device as indicated
herein and a
kit as indicated herein comprising the buprenorphine analogue composition.
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In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) fentanyl or at least one analogue or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
The fentanyl analogue composition will preferably be a preferred composition
as
indicated herein. The content of fentanyl or at least one analogue or
derivative
thereof, such as alfentanil, sulfentanil and remifentanil, will typically be
around 10
to 200 g per hour of depot duration, preferably 25 to 100 g per hour
duration for a
duration of 24 to 170 hours, preferably 48 to 120 (e.g. 72) hours. Fentanyl
analogue
depots may be adininistered by i.m. or s.c. injection, or preferably by
epidural
catheter.
The invention further provides a method of treatment comprising administration
of a
fentanyl analogue composition as described above, especially in a subject in
need
thereof. The method of treatment is particularly for the treatinent of pain,
especially
chronic pain or postoperative pain, where epidural administration may be
preferred.
The invention also provides the use of a fentanyl analogue composition as
described
above in the manufacture of a medicament for the treatment of pain, especially
chronic pain or postoperative pain, where epidural administration may be
preferred.
The invention additionally provides for a pre-filled administration device as
indicated herein and a kit as indicated herein comprising the fentanyl
analogue
composition.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) finasteride or at least one analogue or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
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The finasteride analogue composition will preferably be a preferred
composition as
indicated herein. The content of finasteride will typically be around 10 to
100 mg
per week of depot duration, preferably 15 to 60 mg per week duration for a
duration
of 4 to 24 weeks, preferably 8 to 16 (e.g. 12) weeks.
The invention further provides a method of treatment comprising administration
of a
finasteride analogue composition as described above, especially in a subject
in need
thereof. The method of treatment is particularly for the treatment of male
pattern
baldness and/or enlarged prostate. The invention also provides the use of a
finasteride analogue composition as described above in the manufacture of a
medicament for the treatment of male pattern baldness and/or enlarged
prostate.
The invention additionally provides for a pre-filled administration device as
indicated herein and a kit as indicated herein comprising the finasteride
analogue
composition.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) interferon beta, or at least one analogue or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
The interferon beta analogue composition will preferably be a preferred
composition
as indicated herein. The content of interferon beta or analogue will typically
be
around 0.5 to 10 mg per week of depot duration, preferably 0.7 to 5 mg per
week
duration for a duration of 4 to 24 weeks, preferably 8 to 16 (e.g. 12) weeks.
The invention further provides a method of treatment comprising administration
of a
interferon beta analogue composition as described above, especially in a
subject in
need thereof. The method of treatment is particularly for the treatment of
multiple
sclerosis. The invention also provides the use of a interferon beta analogue
composition as described above in the manufacture of a medicament for the
treatment of multiple sclerosis. The invention additionally provides for a pre-
filled
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administration device as indicated herein and a kit as indicated herein
comprising
the interferon beta analogue composition.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) at least one dopamine agonist;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
The dopamine agonist composition will preferably be a preferred composition as
indicated herein. The content of dopamine agonist, such as pramipexole will
typically be around 1 to 100 mg per week of depot duration, preferably 2 to 50
mg
per week duration for a duration of 4 to 24 weeks, preferably 8 to 16 (e.g.
12)
weeks.
The invention further provides a method of treatment comprising administration
of a
dopamine agonist composition as described above, especially in a subject in
need
thereof. The method of treatment is particularly for the treatment of
parkinson's
disease. The invention also provides the use of a interferon beta analogue
composition as described above in the manufacture of a medicament for the
treatment of parkinson's disease. The invention additionally provides for a
pre-
filled administration device as indicated herein and a kit as indicated herein
comprising the dopamine agonist composition.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) somatotropin, or at least one analogue or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
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The somatotropin analogue composition will preferably be a preferred
composition
as indicated herein. The content of somatotropin or analogue will typically be
around 0.1 to 10 mg per week of depot duration, preferably 0.2 to 8 ing per
week
duration for a duration of 4 to 24 weeks, preferably 8 to 16 (e.g. 12) weeks.
The invention further provides a method of treatment comprising administration
of a
somatotropin analogue composition as described above, especially in a subject
in
need thereof. The method of treatment is particularly for the treatment of
human
growth hormone deficiency and short stature in children and adults. The
invention
also provides the use of a somatotropin analogue composition as described
above in
the manufacture of a medicament for the treatment of human growth hormone
deficiency and short stature in children and adults. The invention
additionally
provides for a pre-filled administration device as indicated herein and a kit
as
indicated herein comprising the somatotropin analogue composition.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) an alpha-agonist such as clonidine, or at least one analogue or derivative
thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
'25
The clonidine analogue composition will preferably be a preferred composition
as
indicated herein. The content of clonidine or analogue will typically around
0.2 to
50 mg per week of depot duration, preferably 0.7 to 25 mg per week duration
for a
duration of 4 to 24 weeks, preferably 8 to 16 (e.g. 12) weeks.
The invention further provides a method of treatment comprising administration
of a
clonidine analogue composition as described above, especially in a subject in
need
thereof. The method of treatment is particularly for the treatment of
hypertension.
The invention also provides the use of a clonidine analogue composition as
described above in the manufacture of a medicament for the treatment of
hypertension. The invention additionally provides for a pre-filled
administration
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device as indicated herein and a kit as indicated herein comprising the
clonidine
analogue composition. The ability to tailor the dose of the depot to
individual
patients is particularly important with alpha-agonist compositions.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) naltrexone, or at least one analogue or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact witli an aqueous fluid.
The naltrexone analogue coinposition will preferably be a preferred
composition as
indicated herein. The content of naltrexone or analogue will typically around
70 to
1000 mg per week of depot duration, preferably 350 to 750 mg per week duration
for a duration of 1 to 6 weeks, preferably 1 to 4 (e.g. 2) weeks.
The invention further provides a method of treatment comprising administration
of a
naltrexone analogue composition as described above, especially in a subject in
need
tliereof. The method of treatment is particularly for the treatment of opoid
addiction
and/or dependence. The invention also provides the use of a naltrexone
analogue
composition as described above in the manufacture of a medicament for the
treatment of opoid addiction and/or dependence. The invention additionally
provides for a pre-filled administration device as indicated herein and a kit
as
indicated herein comprising the naltrexone analogue composition.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) a taxol, or at least one analogue or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
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The taxol analogue composition will preferably be a preferred composition as
indicated herein. The content of taxol or analogue will typically be
paclitaxel or a
structurally related derivative and be present at around 20 to 120 mg per week
of
depot duration, preferably 35 to 80 mg per week duration for a duration of 1
to 12
weeks, preferably 1 to 8 (e.g. 3 or 6) weeks.
The invention further provides a method of treatment comprising administration
of a
taxol analogue composition as described above, especially in a subject in need
thereof. The method of treatment is particularly for the treatment of cancers,
such as
node-positive breast cancer, ovarian cancer, non-small-cell lung cancer and/or
Kaposi's sarcoma. The invention also provides the use of a naltrexone analogue
composition as described above in the manufacture of a medicament for the
treatment of cancers, such as node-positive breast cancer, ovarian cancer, non-
small-
cell lung cancer and/or Kaposi's sarcoma. The invention additionally provides
for a
pre-filled administration device as indicated herein and a kit as indicated
herein
comprising the taxol analogue composition. The ability to tailor the
administered
dose to the patient at the point of delivery is very important in this aspect
of the
invention, since dosages are typically by body surface area.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) bupivacaine or at least one analogue or derivative thereof, such as
levobupivacaine;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
The bupivacaine analogue composition will preferably be a preferred
composition as
indicated herein. The content of bupivacaine or at least one analogue or
derivative
thereof, will typically be around 5 to 200 mg per hour of depot duration,
preferably
10 to 100 mg per hour duration for a duration of 16 to 170 hours, preferably
48 to
120 (e.g. 72) hours. bupivacaine analogue depots may be administered by i.m.
or
s.c. injection, or by epidural catheter or other appropriate route.
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The invention further provides a method of treatment comprising administration
of a
bupivacaine analogue composition as described above, especially in a subject
in
need thereof. The method of treatment is particularly for the treatment of
pain,
especially chronic pain or postoperative pain, where epidural administration
may be
appropriate. The invention also provides the use of a bupivacaine analogue
composition as described above in the manufacture of a medicament for the
treatment of pain, especially chronic pain or postoperative pain, where
epidural
administration may be appropriate. The invention additionally provides for a
pre-
filled administration device as indicated herein and a kit as indicated herein
comprising the bupivacaine analogue composition.
In a still further aspect, the invention also provides a pre-formulation,
comprising a
low viscosity mixture of:
a) at least one diacyl glycerol;
b) at least one phosphatidyl choline;
c) at least one oxygen containing organic solvent;
d) GLP-1 , or at least one analogue, receptor agonist or derivative thereof;
wherein the pre-formulation forms, or is capable of forming, at least one
liquid
crystalline phase structure upon contact with an aqueous fluid.
The GLP-1 analogue composition will preferably be a preferred composition as
indicated herein. The content of GLP-1 or analogue will typically be around
0.05 to
10 mg per week of depot duration, preferably 0.1 to 8 mg per week duration for
a
duration of 1 to 24 weeks, preferably 2 to 16 (e.g. 12) weeks.
Glucagon-like peptide (GLP)-1 is a potent glucoregulatory hormone that is
released
from intestinal L cells into the circulation in response to nutrient ingestion
and
neural and endocrine stimuli. Structurally, GLP-1 is a 37-amino acid peptide
with a
MW of 4,2 KDa, having a sequence highly conserved between different species.
GLP-1 is involved in modification of glucose homeostasis through actions that
include potentiation of glucose-stimulated insulin secretion and biosynthesis
and
suppression of glucagon secretion, gastric emptying, and food intake. The
abilities
of GLP-1 to stimulate insulin secretion and inhibit glucagon release are
glucose-
dependent; thus, the risk of hypoglycemia with GLP-1 administration is low.
GLP-1
also increases beta-cell mass in preclinical models of diabetes through
mechanisms
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that include stimulation of beta-cell proliferation and neogenesis and
inhibition of
beta-cell apoptosis. Studies in both animals and humans indicate that GLP-1
may
also play a protective role in the cardiovascular system.
The combined actions of GLP-1 have generated substantial interest in using
this
peptide as a therapeutic agent for the treatment of type 2 diabetes. However,
the
therapeutic potential of native GLP-1 is limited by its very short plasma half-
life
(below 2 minutes). This is due to both rapid inactivation by the proteolytic
enzyme
dipeptidyl peptidase (DPP)-IV and renal clearance. Consequently, long-acting,
DPP-
IV-resistant GLP-1 analogs have been developed for clinical use, including
exenatide (Byetta; Amylin-Lilly), liraglutide (Novo Nordisk), CJC-1131
(ConjuChem), AVE010 (Zealand Pharma - Sanofi-Aventis), LY548806 (Lilly), and
TH-0318 (TheraTechnologies). All these are once- or twice-daily administration
products; a controlled-release (one week) exentide product (Alkermes-Amylin-
Lilly)
is currently under clinical investigation. These GLP-1 mimetics bind to GLP-1
receptors with similar affinity and produce biological actions identical to
those of
native GLP-1 but are resistant to DPP-IV-mediated inactivation and renal
clearance.
These compounds are able to exert more sustained GLP-1-like activity for
longer
periods of time in vivo. An alternative therapeutic approach for prolonging
the
action of native GLP-1 is to inhibit DPP-IV activity, thereby preventing GLP-1
degradation. Several orally active agents that inhibit DPP-IV activity are
being
evaluated for the treatment of type 2 diabetes.
The invention further provides a method of treatment comprising administration
of a
GLP-1 analogue, composition as described above, especially in a subject in
need
thereof. The method of treatment is particularly for the treatment of
diabetes,
especially type II diabetes. The invention also provides the use of a GLP-1
analogue
composition as described above in the manufacture of a medicament for the
treatment of diabetes, especially type II diabetes. The invention additionally
provides for a pre-filled administration device as indicated, herein and a kit
as
indicated herein comprising the GLP-1 analogue composition.
The Invention will now be further illustrated by reference to the following
non-
limiting Examples and the attached Figures, in which;
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Figure 1 shows leuprolide plasma levels in the rat model following
administration of
leuprolide formulation precursor (0.3 wt% in leuprolide).
Figure 2 shows testosterone plasma levels in the rat model following
administration
of testosterone undecanoate (TEU) and testosterone enanthate (TEE) formulation
precursors (25 wt% in testosterone ester).
Examples:
Example 1: Availability of various liquid crystalline phases in the depot by
choice of composition
Injectable formulations containing different proportions of phosphatidyl
choline
("PC" - Lipoid S 100) and glycerol dioleate (GDO) and with EtOH as solvent
were
prepared to illustrate that various liquid crystalline phases can be accessed
after
equilibrating the depot precursor formulation with excess water.
Appropriate amounts of PC, GDO and EtOH were weighed in glass vials and the
mixture was placed on a shaker until the PC completely dissolved to form a
clear
liquid solution. GDO was then added to form an injectable homogenous solution.
Each formulation was injected in a vial and equilibrated with excess water.
The
phase behaviour was evaluated visually and between crossed polarizes at 25 C.
Results are presented in Table 1.
TABLE 1
Formulation PC (wt%) GDO (wt%) EtOH (wt%) Phase in H20
A 22.5 67.5 10.0 L2
B 28.8 61.2 10.0 I2
C 45.0 45.0 10.0 I2/HI1
D 63.0 27.0 10.0 HII/La,
LZ = reversed micellar phase
12 = reversed cubic liquid crystalline phase
HII = reversed hexagonal liquid crystalline phase
La, = lamellar phase
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Example 2: Viscosity in PC/GDO mixtures on addition of co-solvent
Mixtures of PC/GDO and co-solvent were prepared according to the methods of
Example 1. The EtOH content was adjusted by first evaporating the EtOH from
the
PC/GDO-mixture on a rotary evaporator leaving a viscous liquid mixture of
essentially only PC and GDO. Co-solvents were then added in the proportions
indicated in Table 2 below.
The samples were allowed to equilibrate for several days before viscosity
measurements were performed using a Physica UDS 200 rheometer at 25 C.
TABLE2
Sample PC/GDO EtOH / Glycerol / H20 / Viscosity /
(wt/wt) wt% wt% wt% mPas
1 50/50 3 - - 1900
2 50/50 5 - - 780
3 50/50 7 - - 430
4 50/50 8 - - 300
5 50/50 10 - - 210
6 50/50 15 - - 100
7 45/55 3 - - 1350
8 45/55 5 - - 540
9 45/55 7 - - 320
10 45/55 8 - - 250
11 45/55 10 - - 150
12 45/55 15 - - 85
13 40/60 3 - - 740
14 40/60 5- - - 400
40/60 7 - - 240
16 40/60 8 - - 200
17 40/60 10 - - 130
18 40/60 15 - - 57
19 40/60 - 10 - 8* 10
40/60 - - 3 2.5*108
21 40/60 - - 5 4* 10
This example illustrates the need for a solvent with viscosity lowering
properties in
order to obtain injectable formulations. The mixtures containing glycerol
(sample
19) or water (samples 20 and 21) are too viscous to be injectable at solvent
15 concentrations equivalent to the samples containing EtOH (compare with
samples
13, 14 and 17).
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Example 3: Preparation of depot compositions containing the peptide
leuprolide
Leuprolide acetate is an acetate salt of a synthetic nonapeptide and an
analogue of
gonadotropin-releasing hormone (GnRH) (also known as luteinizing hormone-
releasing hormone (LHRH)). As a result of leuprolide administration to a
subject,
there is an initial increase in follicle stimulating honnone (FSH) and
luteinizing
hormone (LH) secretion- (so-called flare effect) which in turn stimulates the
production of testosterone by the testes in men and estrogens by ovaries in
women.
After about 10 days, a profound hypogonadal effect, equivalent to surgical
castration, is achievQd through down-regulation. Generally this induced and
reversible hypogonadism is the therapeutic goal.
Leuprolide was first mixed with PC, GDO and EtOH where EtOH was added in
excess to dissolve the peptide and the lipids to give a homogenous and clear
solution. Typically, the EtOH content at this stage was about 50-80 wt%. The
excess
EtOH was then removed by rotary evaporation or freeze-drying and the fmal EtOH
content was thereafter adjusted as required. The final compositions of the
samples
are given in Table 3 below.
TABLE 3
Formulation Leuprolide I wt% PC / wt% GDO / wt% EtOH / wt%
A 0.30 47.35 47.35 5.0
B 0.66 47.17 47.17 5.0
C 2.0 46.5 46.5 5.0
D 4.5 45.25 45.25 5.0
E 6.0 44.5 44.5 5.0
Injecting the formulation precursor into excess aqueous phase (syringe 23G;
0.6mm
x 30mm) resulted in a monolithic liquid crystalline phase i.e. leuprolide did
not
change monolith formation and phase behaviour after exposure to an aqueous
environment.
The leuprolide depot precursor formulations in this Example were tested for
stability
against crystallization during storage. Each formulation was stable at 4-8 C
for at
least two weeks.
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Example 4: In vivo release study from depot formulation containing leuprolide
subcutaneously administered
In an in vivo rat model the drug release of leuprolide was followed during 28
days.
The formulation was administered subcutaneously between the scapulae by using
a
syringe (23G, 0.6mm x 25mm). The leuprolide concentration in the rat plasma
was
followed for a period of 28 days (see Figure 1). The dose was 3 mg/kg and the
dose
volume 1 ml/kg corresponding to a drug load of 0.3 wt% leuprolide in the depot
formulation precursor (Formulation A in Example 3).
Figure 1 shows leuprolide plasma levels in the rat model following
administration of
leuprolide formulation precursor (0.3 wt% in leuprolide). It appears that the
investigated formulation gives a release profile with a minimal initial
release (low
"burst") and a sustained release duration of at least 28 days.
Example 5: Preparation of depot compositions containing the peptide goserelin
Goserelin is a potent synthetic decapeptide analogue of luteinizing hormone-
releasing hormone (LHRH), also known as a GnRH agonist analogue. Goserelin
binds to the gonadotropin releasing hormone (GnRH) receptor and, after
prolonged
administration, inhibits endogenous secretion of gonadotropin, resulting in
suppression of sex hormone production in the ovary and testes. This agent
reduces
testosterone production to castration levels and may inhibit androgen receptor-
positive tumor progression.
Goserelin was first mixed with PC, GDO and EtOH where EtOH was added in
excess to dissolve the peptide and the lipids to give a homogenous and clear
solution. The EtOH content at this stage was about 50-80 wt%. The excess EtOH
was then removed by rotary evaporation or freeze-drying and the final EtOH
content
was thereafter adjusted as required. The final compositions of the samples are
given
in Table 4 below.
TABLE 4
Formulation Goserelin / wt% PC / wt% GDO / wt% EtOH / wt%
A 1.08 46.96 46.96 5.00
B 2.16 46.42 46.42 5.00
C 4.32 45.34 45.34 5.00
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Injecting the formulation precursor into excess aqueous phase (syringe 23G;
0.6mm
x 30mm) resulted in a monolithic liquid crystalline phase i.e. goserelin did
not
change monolith formation and phase behaviour after exposure to an aqueous
environment.
The goserelin depot precursor formulations in this Example were tested for
stability
against crystallization during storage. Each formulation was stable at 4-8 C
for at
least two weeks.
Example 6: Preparation of depot compositions containing the peptide
triptorelin
Triptorelin is a potent synthetic decapeptide analogue of luteinizing horinone-
releasing hormone (LHRH), also known as a GnRH agonist analogue. Triptorelizx
binds to the gonadotropin releasing hormone (GnRH) receptor and, after
prolonged
administration, inhibits endogenous secretion of gonadotropin, resulting in
suppression of sex hormone production in the ovary and testes. This agent
reduces
testosterone production to castration levels and may inhibit androgen receptor-
positive tumor progression.
Triptorelin was first mixed with PC, GDO and EtOH where EtOH was added in
excess to dissolve the peptide and the lipids to give a homogenous and clear
solution. The EtOH content at this stage was about 50-80 wt%. The excess EtOH
was then removed by rotary evaporation or freeze-drying and the fmal EtOH
content
was thereafter adjusted as required. The final compositions of the samples are
given
in Table 5 below.
TABLE 5
Formulation Triptorelin / wt% PC / wt% GDO / wt% EtOH / wt%
A 0.75 47.125 47.125 5.00
B 1.5 46.75 46.75 5.00
Injecting the formulation precursor into excess aqueous phase (syringe 23G;
0.6mm
x 30mm) resulted in a monolithic liquid crystalline phase i.e. triptorelin did
not
change monolith formation and phase behaviour after exposure to an aqueous
environment.
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The triptorelin depot precursor formulations in this Example were tested for
stability
against crystallization during storage. Each formulation was stable at 4-8 C
for at
least two weeks.
Example 7: Preparation of depot compositions containing the peptide buserelin
Buserelin is a highly potent synthetic nonapeptide analogue of luteinizing
hormone-
releasing hormone (LHRH), also known as a GnRH agonist analogue. Buserelin
binds to the gonadotropin releasing hormone (GnRH) receptor and, after
prolonged
administration, inhibits endogenous secretion of gonadotropin, resulting in
suppression of sex ltormone production in the ovary and testes. This agent
reduces
testosterone production to castration levels and may inhibit androgen receptor-
positive tumor progression.
Buserelin was first mixed with PC, GDO and EtOH where EtOH was added in
excess to dissolve the peptide and the lipids to give a homogenous and clear
solution. The EtOH content at this stage was about 50-80 wt%. The excess EtOH
was then removed by rotary evaporation or freeze-drying and the final EtOH
content
was thereafter adjusted as required. The final compositions of the samples are
given
in Table 6 below.
TABLE 6
Formulation Buserelin / wt% PC / wt% GDO / wt% EtOH / wt%
A 0.66 47.17 47.17 5.00
B 1.32 46.84 46.84 5.00
C 1.98 46.51 46.51 5.00
Injecting the formulation precursor into excess aqueous phase (syringe 23G;
0.6mm
x 30mm) resulted in a monolithic liquid crystalline phase i.e. buserelin did
not
change monolith formation and phase behaviour after exposure to an aqueous
environment.
The buserelin depot precursor formulations in this Example were tested for
stability
against crystallization during storage. Each formulation was stable at 4-8 C
for at
least two weeks.
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Example 8: Degradation of depot formulation in the rat
Various volumes (1, 2, 6 ml/kg) of the depot precursor (36%wt PC, 54%wt GDO,
and 10%wt EtOH) were injected in the rat and were removed again after a period
of
14 days. It was found that substantial amounts of the formulations were still
present
subcutaneously in the rat after this time, see Table 7.
TABLE 7
Dose (ml/kg) Mean diameter day 3 (mm) Mean diameter day 14 (mm)
1 (n=3) 15.8 12.5
2 (n=3) 18.5 15.3
6 (n=3) 23.3 19.3
Example 8B: Further compositions containing GnRH agonist analogues
Formulations were prepared as described in Examples 3, 5, 6 and 7 by mixing
the
peptide active with a mixture of GDO (at one of several purity levels), PC,
ethanol
and optionally dioleoyl PG in the proportions (by weight) indicated in Table
8.
TABLE 8. Abbreviations: LEU = Leuprolide; GOS = Goserelin; T.RI = Triptorelin;
BUS = Buserelin
Formulation Peptide EtOH PC GDOI GDO2 GDO3 DOPG
1 2.25 10 35.1 - - 52.65 -
(LEU)
2 2.25 10 35.1 52.65 - - -
(LEU)
3 2.25 10 35.1 - 52.65 - -
(LEU)
4 2.25 7 36.3 - - 54.45 -
(LEU)
5 2.16 10 35.14 - - 52.70 -
(GOS)
6 2.16 7 36.34 - - 54.50 -
(GOS)
7 2.16 5 37.14 - - 55.70 -
(GOS)
8 1.50 10 35.4 - - 53.1 -
(TRI)
9 1.50 7 36.6 - - 54.9 -
(TRI)
10 1.50 5 37.4 - - 56.1 -
(TRI)
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11 1.32 10 35.47 - - 53.21 -
(BUS)
12 1.32 7 36.67 - - 55.01 -
(BUS)
13 1.32 5 37.47 - - 56.21 -
(BUS)
14 2.25 10 39.49 - - 48.26 -
(LEU)
15 2.25 5 45.375 - - 45.375 2
(LEU)
16 2.25 5 44.375 - - 44.375 4
(LEU)
where EtOH is ethanol,,PC is LIPOID S 100 soybean phosphatidylcholine, GDO is
glycerol dioleate
and DOPG is dioleoyl phosphatidylglycerol
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO1 10.9% 87.5% 1.4%
GDO2 4.2% 92.1% 3.5%
GDO3 0.5% 95.3% 4.0%
Example 8C: Preparation of depot compositions of glucagon-like peptide 1
(GLP-1)
Depot precursors of GLP-1 were prepared in two different ways:
1) GLP-1 was first mixed with PC, GDO and EtOH where EtOH was added in
excess to facilitate mixing. Typically, the EtOH content at this stage was
about 50-80 wt%. The excess EtOH was then removed by rotary evaporation
or freeze-drying and the final EtOH content was thereafter adjusted as
required.
2) GLP-1 was first dissolved in a small amount of sterile water. A pre-made
liquid mixture of PC, GDO and EtOH, where the EtOH content was about 5-
10% by weight, was then added to the GLP-1/water solution. The resulting
mixture was mixed by vortex mixing for 1 min.
The final compositions of the samples are given in Table 9 below. Several
purity
levels of GDO and both soy and egg phosphatidylcholine (PC) were used.
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TABLE 9: Coinpositions containing GLP-1
Formulation GLP-1 PC / GDO1 / GDO2 / GDO3 / EtOH / H20 /
/ Wt% Wt% Wt% Wt% Wt% Wt% Wt%
A 0.5 44.75 44.75 - - 10 -
B 0.5 44.75 - 44.75 - 10 -
C 0.5 44.75 - - 44.75 10 -
D 1.0 44.5 - - 44.5 10 -
E 1.0 46 - - 46 7 -
F 1.0 47 - - 47 5 -
G 2.0 44 - - 44 10 -
H 2.0 45.5 - - 45.5 7 -
I 2.0 46.5 - - 46.5 5 -
J 3.0 46 - - 46 5 -
K 0.5 35.775 - - 43.725 10 10
L 1.0 35.55 - - 43.45 10 10
M 2.0 37.35 - - 45.65 5 10
N 2.0 32.85 - - 40.15 10 15
0 2.0 30.4 - - 45.6 10 12
P 3.0 30 - - 45 10 12
Q 3.0 31.875 - - 43.125 10 12
R 3.0 32.4 - - 39.6 10 15
S 2.0* 46.5 - - 46.5 5 -
T 2.0* 32.85 - - 40.15 10 15
U 2.0* 30.4 - - 45.6 10 12
where EtOH is ethanol, PC is LIPOID S 100 soybean phosphatidylcholine or
LIPOID E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate
TABLE 10: GDO qualities used
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO1 10.9% 87.5% 1.4%
GDO2 4.2% 92.1% 3.5%
GDO3 0.5% 95.3% 4.0%
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Example 9: Preparation of depot compositions of paclitaxel
Depot precursors of paclitaxel were prepared by mixing paclitaxel, PC, GDO and
EtOH where EtOH was added in excess to facilitate mixing. Typically, the EtOH
content at this stage was about 50-80 wt%. The excess EtOH was then removed by
rotary evaporation or freeze-drying and the final EtOH content was thereafter
adjusted as required.
The final compositions of the samples are given in Table 11 below. Both soy
and
egg phosphatidylcholine (PC) were used in the compositions.
TABLE 11: Compositions containing paclitaxel
Formulation Pacl'itaxel / wt% PC / wt% GDO3 / wt% EtOH / wt%
A 0.5 44.75 44.75 10
B 0.5 46.25 46.25 7
C 0.5 47.25 47.25 5
D 0.5 37.8 56.7 5
E 0.5* 47.25 47.25 5
F 1.0 44.5 44.5 10
G 1.0 46 46 7
H 1.0 47 47 5
I 1.0 37.6 56.4 5
J 1.0* 47 47 5
K 2.0 44 44 10
L 2.0 45.5 45.5 7
M 2.0 46.5 46.5 5
N 2.0 37.2 55.8 5
0 2.0* 46.5 46.5 5
P 5 42.5 42.5 10
Q 5 44 44 7
R 5 45 45 5
S 5 36 54 5
T 5* 45 45 5
U 10 40 40 10
V 10 41.5 41.5 7
W 10 42.5 42.5 5
X 10 34 51 5
Y 10* 42.5 42.5 5
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where EtOH is ethanol, PC is LIPOID S100 soybean phosphatidylcholine or LIPOID
E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 10: Preparation of depot compositions of interferon beta lA
Depot precursors of interferon beta 1 A were prepared in two different ways:
1) Interferon b6ta 1A was first mixed with PC, GDO and EtOH where EtOH
was added in excess to facilitate mixing. Typically, the EtOH content at this
stage was about 50-80 wt%. The excess EtOH was then removed by rotary
evaporation or freeze-drying and the final EtOH content was thereafter
adjusted as required.
2) Interferon beta 1 A was first dissolved in a small amount of sterile water.
A
pre-made liquid mixture of PC, GDO and EtOH, where the EtOH content
was about 5-10% by weight, was then added to the Interferon beta lA/water
solution. The resulting mixture was mixed by vortex mixing for 1 min.
The final compositions of the samples are given in Table 12 below. Both soy
and
egg phosphatidylcholine (PC) were used.
TABLE 12: Compositions containing interferon beta 1 A
/
Formulation Interferon beta 1A / PC / GDO3 / EtOH / H20
wt% wt% wt% wt% wt%
A 0.03 44.985 44.985 10 -
B 0.03 46.485 46.485 7 -
C 0.03 47.485 47.485 5 -
D 0.05 44.975 44.975 10 -
E 0.05 46.475 46.475 7 -
F 0.05 47.475 47.475 5 -
G 0.1 37.96 56.94 5 -
H 0.1 47.45 47.45 5 -
I 0.1* 37.96 56.94 5 -
J 0.1* 47.45 47.45 5 -
K 0.05 35.98 43.97 10 10
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L 0.05 37.98 45.97 10 10
M 0.05 31.98 47.97 10 10
N 0.1 35.96 43.94 10 10
0 0.1 31.96 47.94 10 10
P 0.1* 35.96 43.94 10 10
Q 0.1* 31.96 47.94 10 10
R 0.2 33.9 45.9 10 10
S 0.2 35.9 43.9 10 10
where EtOH is ethanol, PC is LIPOID S100 soybean phosphatidylcholine or LIPOID
E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 11: Preparation of depot compositions of human growth hormone
(hGH)
Depot precursors of hGH were prepared in two different ways:
1) hGH was first mixed with PC, GDO and EtOH where EtOH was added in
excess to facilitate mixing. Typically, the EtOH content at this stage was
about 50-80 wt%. The excess EtOH was then removed by rotary evaporation
or freeze-drying and the final EtOH content was thereafter adjusted as
required.
2) hGH was first mixed in a small amount of sterile water. A pre-made liquid
mixture of PC, GDO and EtOH, where the EtOH content was about 5-10%
by weight, was then added to the hGH/water mixture. The resulting mixture
was mixed by vortex rnixing for 1 min.
The final compositions of the samples are given in Table 13 below. Both soy
and
egg phosphatidylcholine (PC) were used.
TABLE 13 Compositions containing hGH
Formulation hGH / wt% PC / wt% GDO3 / wt% EtOH / wt% H20 / wt%
A 0.5 44.75 44.75 10 -
B 0.5 46.25 46.25 7 -
C 0.5 47.25 47.25 5 -
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D 0.5 33.53 40.97 10 15
E 0.5 35.78 43.72 10 10
F 0.5 37.13 45.37 7 10
G 1 47 47 5 -
H 1 31.6 47.4 10 10
I 1 34.65 42.35 10 12
J 1 33.75 41.25 10 14
K 1 33.3 40.7 10 15
L 1* 34.65 42.35 10 12
M 1'.2 34.68 39.12 10 15
N 1.2 33.21 40.59 10 15
0 1.2* 34.68 39.12 10 15
P 1.2* 33.21 40.59 10 15
where EtOH is ethanol, PC is LIPOID S 100 soybean phosphatidylcholine or
LIPOID E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 12: Preparation of depot compositions of naltrexone
Depot precursors of naltrexone were prepared by mixing naltrexone, PC, GDO and
EtOH where EtOH was added in excess to facilitate mixing. Typically, the EtOH
content at this stage was about 50-80 wt%. The excess EtOH was then removed by
rotary evaporation or freeze-drying and the final EtOH content was thereafter
adjusted as required.
The fmal compositions of the samples are given in Table 14 below.
TABLE 14: Compositions containing naltrexone
Formulation Naltrexone / wt% PC / wt% GDO3 / wt% EtOH / wt%
A 5 42.5 42.5 10
B 5 44 44 7
C 5 45 45 5
D 5 36 54 5
E 5* 45 45 5
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F 10 40 40 10
G 10 41.5 41.5 7
H 10 42.5 42.5 5
I 10 34 51 5
J 10* 42.5 42.5 5
K 15 37.5 37.5 10
L 15 39 39 7
M 15 40 40 5
N 15 32 48 5
0 ~ 15 * 40 40 5
P 20 35 35 10
Q 20 36.5 36.5 7
R 20 37.5 37.5 5
S 20 30 45 5
T 20* 37.5 37.5 5
where EtOH is ethanol, PC is LIPOID S100 soybean phosphatidylcholine or LIPOID
E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 13: Preparation of depot compositions of bupivacaine and
levobupivacaine
Depot precursors of bupivacaine (or levobupivacaine) were prepared by mixing
bupivacaine or levobupivacaine, PC, GDO and EtOH where EtOH was added in
excess to facilitate mixing. Typically, the EtOH content at this stage was
about 50-
80 wt%. The excess EtOH was then removed by rotary evaporation or freeze-
drying
and the final EtOH content was thereafter adjusted as required.
The final compositions of the samples are given in Table 15 below. Both soy
and
egg phosphatidylcholine (PC) were used in the compositions.
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TABLE 15: Compositions containing bupivacaine or levobupivacaine
Formulation Bupivacaine or PC / GDO3 I EtOH /
levobupivacaine wt% wt% wt%
/ wt%
A 5 42.5 42.5 10
B 5 44 44 7
C 5 45 45 5
D 5 36 54 5
E 5* 45 45 5
F 10 40 40 10
G 10 41.5 41.5 7
H 10 42.5 42.5 5
I 10 34 51 5
J 10* 42.5 42.5 5
K 15 37.5 37.5 10
L 15 39 39 7
M 15 40 40 5
N 15 32 48 5
0 15* 40 40 5
where EtOH is ethanol, PC is LIPOID S100 soybean phosphatidylcholine or LIPOID
E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 14: Preparation of depot compositions of pramipexole
Depot precursors of pramipexole were prepared by mixing pramipexole, PC, GDO
and EtOH where EtOH was added in excess to facilitate mixing. Typically, the
EtOH content at this stage was about 50-80 wt%. The excess EtOH was then
removed by rotary evaporation or freeze-drying and the final EtOH content was
thereafter adjusted as required.
The final compositions of the samples are given in Table 16 below. Both soy
and
egg phosphatidylcholine (PC) were used in the compositions.
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TABLE 16: Compositions containing pramipexole
Formulation Pramipexole / wt% PC / wt% GDO3 / wt% EtOH / wt%
A 1.0 44.5 44.5 10
B 1.0 46 46 7
C 1.0 47 47 5
D 1.0 37.6 56.4 5
E 1.0* 47 47 5
F 3 43.5 43.5 10
G 3 45 45 7
H 3 46 46 5
I 3 36.8 55.2 5
J 3* 46 46 5
K 5 42.5 42.5 10
L 5 44 44 7
M 5 45 45 5
N 5 36 54 5
0 5* 45 45 5
where EtOH is ethanol, PC is LIPOID S100 soybean phosphatidylcholine or LIPOID
E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 15: Preparation of depot compositions of clonidine
Depot precursors of clonidine were prepared by mixing clonidine, PC, GDO and
EtOH where EtOH was added in excess to facilitate mixing. Typically, the EtOH
content at this stage was about 50-80 wt%. The excess EtOH was then removed by
rotary evaporation or freeze-drying and the final EtOH content was thereafter
adjusted as required.
The final compositions of the samples are given in Table 17 below. Both soy
and
egg phosphatidylcholine (PC) were used in the compositions.
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TABLE 17: Compositions containing clonidine
Formulation Clonidine / wt% PC / wt% GDO3 / wt% EtOH / wt%
A 1.0 44.5 44.5 10
B 1.0 46 46 7
C 1.0 47 47 5
D 1.0 37.6 56.4 5
E 1.0* 47 47 5
F 3 43.5 43.5 10
G 3 45 45 7
H ; 3 46 46 5
I 3 36.8 55.2 5
J 3* 46 46 5
K 5 42.5 42.5 10
L 5 44 44 7
M 5 45 45 5
N 5 36 54 5
0 5* 45 45 5
where EtOH is ethanol, PC is LIPOID S100 soybean phosphatidylcholine or LIPOID
E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 16: Preparation of depot compositions of levothyroxine
Depot precursors of levothyroxine were prepared by mixing levothyroxine, PC,
GDO and EtOH where EtOH was added in excess to facilitate mixing. Typically,
the
EtOH content at this stage was about 50-80 wt%. The excess EtOH was then
removed by rotary evaporation or freeze-drying and the final EtOH content was
thereafter adjusted as required.
The final compositions of the samples are given in Table 18 below. Both soy
and
egg phosphatidylcholine (PC) were used in the compositions.
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TABLE 18: Compositions containing levothyroxine
Formulation Levothyroxine / wt% PC / wt% GDO3 / wt% EtOH / wt%
A 0.5 44.75 44.75 10
B 0.5 46.25 46.25 7
C 0.5 47.25 47.25 5
D 0.5 37.8 56.7 5
E 0.5* 47.25 47.25 5
F 1.5 44.25 44.25 10
G 1.5 45.75 45.75 7
H ' 1.5 46.75 46.75 5
I 1.5 37.4 56.1 5
J 1.5* 46.75 46.75 5
K 3 43.5 43.5 10
L 3 45 45 7
M 3 46 46 5
N 3 36.8 55.2 5
0 3* 46 46 5
where EtOH is ethanol, PC is LIPOID S 100 soybean phosphatidylcholine or
LIPOID E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 17: Preparation of depot compositions of buprenorphine
Depot precursors of buprenorphine were prepared by mixing buprenorphine, PC,
GDO and EtOH where EtOH was added in excess to facilitate mixing. Typically,
the
EtOH content at this stage was about 50-80 wt%. The excess EtOH was then
removed by rotary evaporation or freeze-drying and the final EtOH content was
thereafter adjusted as required.
The final compositions of the samples are given in Table 19 below. Both soy
and
egg phosphatidylcholine (PC) were used in the compositions.
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TABLE 19: Compositions containing buprenorphine
Formulation Buprenorphine / wt% PC / wt /a GDO3 / wt% EtOH I wt%
A 1.0 44.5 44.5 10
B 1.0 46 46 7
C 1.0 47 47 5
D 1.0 37.6 56.4 5
E 1'.0* 47 47 5
F 3 43.5 43.5 10
G 3 45 45 7
H 3 46 46 5
I 3 36.8 55.2 5
J 3* 46 46 5
K 5 42.5 42.5 10
L 5 44 44 7
M 5 45 45 5
N 5 36 54 5
0 5* 45 45 5
where EtOH is ethanol, PC is LIPOID S 100 soybean phosphatidylcholine or
LIPOID E 80 egg
phosphatidyicholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 18: Preparation of depot compositions of testosterone esters
Depot precursors of the undecanoate and enanthate esters of testosterone were
prepared by mixing the testosterone esters, PC, GDO and EtOH, where EtOH was
added in excess to facilitate mixing. Typically, the EtOH content at this
stage was
about 50-80 wt%. The excess EtOH was then removed by rotary evaporation or
freeze-drying and the final EtOH content was thereafter adjusted as required.
The final compositions of the samples are given in Table 20 below. Both soy
and
egg phosphatidylcholine (PC) were used in the compositions.
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TABLE 20: Compositions containing testosterone undecanoate or testosterone
enanthate
Formulation Testosterone undecanoate or PC / GDO3 / EtOH /
testosterone enanthate / wt% wt% wt% wt%
A 10 40 40 10
B 10 41.5 41.5 7
C 10 42.5 42.5 5
D 10 34 51 5
E 10* 42.5 42.5 5
F 15 37.5 37.5 10
G 15 39 39 7
H 15 40 40 5
I 15 32 48 5
J 15* 40 40 5
K 20 35 35 10
L 20 36.5 36.5 7
M 20 37.5 37.5 5
N 20 30 45 5
0 20* 37.5 37.5 5
P 25 32.5 32.5 10
Q 25 34 34 7
R 25 35 35 5
S 25 28 42 5
T 25* 35 35 5
where EtOH is ethanol, PC is LIPOID S100 soybean phosphatidylcholine or LIPOID
E 80 egg
phosphatidylcholine (marked with *) and GDO is glycerol dioleate (see below)
GDO quality Monoglycerides Diglycerides Triglycerides
(according to AC)
GDO3 0.5% 95.3% 4.0%
Example 19: In vivo release study from depot formulations containing
testosterone esters subcutaneously administered
In an in vivo rat model the drug release of testosterone undecanoate and
testosterone
enanthate was followed during 28 days. The formulations were administered
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subcutaneously between the scapulae by using a syringe (23G, 0.6mm x 25mm).
The testosterone concentration in the rat plasma was followed for a period of
28
days (see Figure 2). The dose was 125 mg/kg and the dose volume 0.5 ml/kg
corresponding to a drug load of 25 wt% testosterone ester in the depot
formulation
precursor (Formulation R in Example 18).
Figure 2 shows testosterone plasma levels in the rat model following
administration
of testosterone undecanoate (TEU) and testosterone enanthate (TEE) formulation
precursors (25 wt% in testosterone ester). It appears that the investigated
forinulations give release profiles with a sustained release duration of at
least 28
days.
Example 20: Preparation of a depot precursor of hGH using a 2-part mixing
device
A depot precursor of hGH was prepared by first dissolving 7.5 mg of hGH in
0.15 g
of sterile water. This solution was withdrawn into a 1 mL glass syringe. A
liquid
solution containing PC, GDO and EtOH (PC/GDO/EtOH = 40.5/49.5/10 wt%) (0.84
g) was withdrawn into a second glass syringe.
The two syringes containing hGH/water and the lipid mixture, respectively,
were
connected using a female to female Luer adapter and the two solutions were
mixed
by repeatedly pushing the content back and forth. After about 15 cycles of
pushing
back and forth, the depot precursor was collected in one of the syringes and
injected
into saline using a 23 gauge needfe.
Example 21: Preparation of a depot precursor of interferon beta lA using a 2-
part mixing device
A depot precursor of interferon beta 1A was prepared by first dissolving 1.0
mg of
interferon beta lA in 0.1 g of sterile water. This solution was withdrawn into
a 1 mL
glass syringe. A liquid solution containing PC, GDO and EtOH (PC/GDO/EtOH =
40.5/49.5/10 wt%) (0.9 g) was withdrawn into a second glass syringe.
The two syringes containing interferon beta lA/water and the lipid mixture,
respectively, were connected using a female to female Luer adapter and the two
solutions were mixed by repeatedly pushing the content back and forth. After
about
15 cycles of pushing back and forth, the depot precursor was collected in one
of the
syringes and injected into saline using a 23 gauge needle.
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Legend to Figures:
Figure 1: Leuprolide plasma levels in the rat model following administration
of
leuprolide (3 mg/kg) formulation precursor (0.3% by weight leuprolide)
according
to Example 4.
Figure 2: Testosterone plasma levels in the rat model following administration
of
different testosterone derivatives according to Exainple 19 (TEU =
testosterone
undecanoate and TEE = testosterone enanthate).
